1. Field of the Invention
The invention relates to a method of extracting all metabolites existing in a biological sample to detect a comprehensive difference between metabolites of a control group and a test group, thereby verifying significance thereof. The biological sample being analyzed by the invention includes all materials existing in an organism, such as blood, blood plasma, serum, urine, feces, cerebrospinal fluid, tissue, hair, serous fluid, gastric juice and the like.
2. Description of the Related Art
Until now, a research using metabolite is mainly concentrated on a targeted metabolomics research of quantitatively analyzing a reference standard of a specific metabolome in a cell under disease state or gene mutated state and then comparing changes in an amount or concentration of metabolites of a test group and a control group or a ratio of amounts or concentrations of metabolites related to a metabolic pathway, so as to clarify a defect occurring in the metabolic pathway when a disease or gene mutation occurs and to induce a disease diagnosis biomarker using it.
However, there are various metabolites in the organism and it is very difficult in a test to acquire a reference standard of all the metabolites and to conduct a quantitative analysis of it. Accordingly, it has appeared a global metabolomics research as a non-targeted metabolomics, which is capable of verifying a change of the metabolites in the organism on the whole without the complicated analysis steps.
Actually, such research has been much conducted for a bacteria or plant in which the number of the metabolites existing in the organism is relatively small [Phytochemistry, 62, 929-937 (2003); Phytochemistry, 62, 817-836 (2003); Phytochemistry, 62, 987-995 (2003); Journal of Integrative Biology, 6, 217-234 (2002); Electrophoresis, 23, 1642-1651 (2002); Nature Biotechnology, 18, 1157-1161 (2000); Anal. Chem., 76, 619-626 (2004); Plant Cell, 14, 1437-1440 (2002); Advan. Enzyme Regul. 43, 67-76 (2003); International Immunopharmacology, 4, 1499-1514 (2004)]. However, regarding all the animal samples including human, an extract method and an analysis are complicated, there are differences between the individual organisms and an analysis method which is an indicator does not exist, so that the above research has not been almost applied.
Therefore, it is seriously needed to standardize a method of extracting a biological sample and to develop a method of connecting a device analysis and statistics enabling a metabolite profiling of a test group and a control group, which is obtained by a device analysis chromatogram, to be statistically analyzed.